gateway bp clonase ii enzyme mix  (Thermo Fisher)


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    Name:
    Gateway BP Clonase Enzyme Mix
    Description:
    Gateway BP Clonase enzyme contains both Int Integrase and IHF Integration Host Factor proteins that catalyze the in vitro recombination of PCR products or DNA segments from clones containing attB sites and a Donor vector containing attP sites to generate Entry clones BP Clonase II enzyme format is a single mix of both enzyme and buffer to ensure enzyme stability and enable convenient 10µl reaction set up with fewer pipetting steps which both maximize recombination efficiency and minimize pippetting The original format of BP Clonase enzyme is the stand alone enzyme blend and reaction buffer provided in separate tubes
    Catalog Number:
    11789013
    Price:
    None
    Applications:
    BP Clonase®|Cloning|Gateway Recombination|Gateway Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher gateway bp clonase ii enzyme mix
    Gateway® BP and LR reactions. BP and LR <t>Clonase®</t> facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.
    Gateway BP Clonase enzyme contains both Int Integrase and IHF Integration Host Factor proteins that catalyze the in vitro recombination of PCR products or DNA segments from clones containing attB sites and a Donor vector containing attP sites to generate Entry clones BP Clonase II enzyme format is a single mix of both enzyme and buffer to ensure enzyme stability and enable convenient 10µl reaction set up with fewer pipetting steps which both maximize recombination efficiency and minimize pippetting The original format of BP Clonase enzyme is the stand alone enzyme blend and reaction buffer provided in separate tubes
    https://www.bioz.com/result/gateway bp clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 247 article reviews
    Price from $9.99 to $1999.99
    gateway bp clonase ii enzyme mix - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes"

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

    Journal: Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.]

    doi: 10.1002/0471142727.mb0320s110

    Gateway® BP and LR reactions. BP and LR Clonase® facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.
    Figure Legend Snippet: Gateway® BP and LR reactions. BP and LR Clonase® facilitate the recombination between att B and att P and between att L and att R, generating att L and att B sequences, respectively.

    Techniques Used:

    2) Product Images from "Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT)"

    Article Title: Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT)

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0144922

    Map of pGADT7-DEST. This plasmid was modified from pGADT7-Rec (Clontech Laboratories); the SMART III and CDS III sequences were replaced by attR1 and attR2, respectively. Following recombination catalysis using the Gateway ® LR Clonase ™ II enzyme mix (Invitrogen), attR1-Cm-ccdB-attR2 was replaced by attB1-cDNA insert-attB2.
    Figure Legend Snippet: Map of pGADT7-DEST. This plasmid was modified from pGADT7-Rec (Clontech Laboratories); the SMART III and CDS III sequences were replaced by attR1 and attR2, respectively. Following recombination catalysis using the Gateway ® LR Clonase ™ II enzyme mix (Invitrogen), attR1-Cm-ccdB-attR2 was replaced by attB1-cDNA insert-attB2.

    Techniques Used: Plasmid Preparation, Modification

    3) Product Images from "A novel process of viral vector barcoding and library preparation enables high-diversity library generation and recombination-free paired-end sequencing"

    Article Title: A novel process of viral vector barcoding and library preparation enables high-diversity library generation and recombination-free paired-end sequencing

    Journal: Scientific Reports

    doi: 10.1038/srep37563

    Process for efficient, zero-background, cloning of uniquely barcoded dA-tailed fragment libraries. (a) To achieve ligation of one unique fragment into each plasmid backbone, a relatively small cloning vector (5.7 kb) was generated containing a ccdB toxin gene flanked by two specifically tailored XcmI restriction enzyme cleavage sites. (b) The sequence is tailored so that XcmI enzyme digestion leaves a single 5′ dT-overhang on both open ends of the backbone generating a T-vector where the dA-tailed DNA fragments can be efficiently ligated without directional bias. (c) The cloning vector can have a flexible design with any 5′ and 3′ domain modulated by the inserted fragment. To allow for unique barcoding of each fragment together with the surrounding 5′ and 3′ domains, the region of interest in the cloning vector is then exposed to a two-step PCR amplification where a 5′ AttB1 site is inserted in the first amplification (20 cycles amplification). (d) The barcode together with the AttB2 site are added through a PCR with only a single elongation step, ensuring that each unique barcode is utilized only once and is not transferred to other amplicons due to PCR template switching. (e) The library of uniquely barcoded PCR amplicons is then inserted into the viral vector backbone using the “Gateway” BP clonase recombination reaction. (f) Chemically or electro-competent bacteria are then transformed and a small fraction of the transformation reaction plated for a rough estimation of total number of colonies. Generation of empty backbones ( i.e., missing a genomic fragment) is kept to an absolute minimal as any such plasmid would contain the ccdB toxin gene providing negative selection pressure.
    Figure Legend Snippet: Process for efficient, zero-background, cloning of uniquely barcoded dA-tailed fragment libraries. (a) To achieve ligation of one unique fragment into each plasmid backbone, a relatively small cloning vector (5.7 kb) was generated containing a ccdB toxin gene flanked by two specifically tailored XcmI restriction enzyme cleavage sites. (b) The sequence is tailored so that XcmI enzyme digestion leaves a single 5′ dT-overhang on both open ends of the backbone generating a T-vector where the dA-tailed DNA fragments can be efficiently ligated without directional bias. (c) The cloning vector can have a flexible design with any 5′ and 3′ domain modulated by the inserted fragment. To allow for unique barcoding of each fragment together with the surrounding 5′ and 3′ domains, the region of interest in the cloning vector is then exposed to a two-step PCR amplification where a 5′ AttB1 site is inserted in the first amplification (20 cycles amplification). (d) The barcode together with the AttB2 site are added through a PCR with only a single elongation step, ensuring that each unique barcode is utilized only once and is not transferred to other amplicons due to PCR template switching. (e) The library of uniquely barcoded PCR amplicons is then inserted into the viral vector backbone using the “Gateway” BP clonase recombination reaction. (f) Chemically or electro-competent bacteria are then transformed and a small fraction of the transformation reaction plated for a rough estimation of total number of colonies. Generation of empty backbones ( i.e., missing a genomic fragment) is kept to an absolute minimal as any such plasmid would contain the ccdB toxin gene providing negative selection pressure.

    Techniques Used: Clone Assay, Ligation, Plasmid Preparation, Generated, Sequencing, Polymerase Chain Reaction, Amplification, Transformation Assay, Selection

    Related Articles

    DNA Extraction:

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes
    Article Snippet: .. 150 ng/µl entry vector (e.g., pDONR™221; Life Technologies) Gateway® BP Clonase® II enzyme mix (Life Technologies) 10 to 50 ng/µl att B-PCR product prepared according to the Gateway® scheme (Basic Protocol 1) PCR-grade H2 O LB plates supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) LB medium supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) 50% (v/v) glycerol, sterile Plasmid DNA isolation kit (e.g., Qiagen; optional) 96-well plate or 1.5 ml microcentrifuge tubes Multichannel pipettor 25°C incubator Additional reagents and equipment for growth of T1 phage resistant (T1R ) E. coli competent cells (e.g. One Shot® MAX Efficiency® DH5α™ competent cells; Life Technologies) ( unit 1.8 ), DNA miniprep ( unit 1.6 or commercial DNA prep kit), and DNA sequencing (Chapter 7). ..

    Clone Assay:

    Article Title: Metabolic Characterization of the Anthocyanidin Reductase Pathway Involved in the Biosynthesis of Flavan-3-ols in Elite Shuchazao Tea (Camellia sinensis) Cultivar in the Field
    Article Snippet: .. The resulting PCR products were purified and then cloned to the entry vector pDONR207 with Gateway® BP Clonase® Enzyme mix according to the manufacturer’s instructions (Invitrogen). .. The resulting ligation products were introduced to competent DH5α cells, which were selected on agar-solidified LB plates containing gentamycin.

    Article Title: Cellular diversity within embryonic stem cells: pluripotent clonal sublines show distinct differentiation potential
    Article Snippet: .. Lentivectors and ESC transduction To generate entry vectors, the promoters [βIII-tubulin promoter (βIIIp) and Tα-1] and genes of interest (GFP and H2B-mRFP1) were cloned into pDONRP4-P1R and pDONR221, respectively, using the Gateway® BP clonase enzyme mix (Invitrogen). .. The resulting entry vectors were then recombined into 2K7bsd or 2K7neo lentivectors using the Gateway® LR plus clonase enzyme mix (Invitrogen).

    Article Title: A novel process of viral vector barcoding and library preparation enables high-diversity library generation and recombination-free paired-end sequencing
    Article Snippet: .. The cloning was performed through a recombination reaction using Gateway BP Clonase II Enzyme Mix (Life Technologies). .. 150 ng of vector was used with 100 ng of the barcoded PCR-product from the previous step, according to manufacturer’s protocol.

    Article Title: PhoPR Contributes to Staphylococcus aureus Growth during Phosphate Starvation and Pathogenesis in an Environment-Specific Manner
    Article Snippet: .. Fragments were cloned into the pKOR1 knockout vector via site-specific recombination using the Gateway BP Clonase II enzyme mix (Thermo Fisher Scientific). .. The deletions were constructed via allelic exchange in Newman and the Δ pstSCAB Δ nptA mutant ( ) , as described previously ( ).

    Transduction:

    Article Title: Cellular diversity within embryonic stem cells: pluripotent clonal sublines show distinct differentiation potential
    Article Snippet: .. Lentivectors and ESC transduction To generate entry vectors, the promoters [βIII-tubulin promoter (βIIIp) and Tα-1] and genes of interest (GFP and H2B-mRFP1) were cloned into pDONRP4-P1R and pDONR221, respectively, using the Gateway® BP clonase enzyme mix (Invitrogen). .. The resulting entry vectors were then recombined into 2K7bsd or 2K7neo lentivectors using the Gateway® LR plus clonase enzyme mix (Invitrogen).

    Construct:

    Article Title: A Staphylococcus aureus TIR-domain protein virulence factor blocks TLR2-mediated NF-κB signaling
    Article Snippet: .. The fusion construct was subcloned into the temperature sensitive pKOR1 ( ) using Gateway BP clonase II enzyme mix (Invitrogen, USA). .. The BP reaction product was transformed into DC10b ultra competent cells.

    Purification:

    Article Title: Metabolic Characterization of the Anthocyanidin Reductase Pathway Involved in the Biosynthesis of Flavan-3-ols in Elite Shuchazao Tea (Camellia sinensis) Cultivar in the Field
    Article Snippet: .. The resulting PCR products were purified and then cloned to the entry vector pDONR207 with Gateway® BP Clonase® Enzyme mix according to the manufacturer’s instructions (Invitrogen). .. The resulting ligation products were introduced to competent DH5α cells, which were selected on agar-solidified LB plates containing gentamycin.

    Article Title: Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT)
    Article Snippet: .. Purified cDNA ligated with three different forward adapters were mixed and recombined with the pDONR™ 222 plasmid vector via catalysis, using the Gateway® BP Clonase™ II Enzyme Mix (Invitrogen). ..

    Knock-Out:

    Article Title: PhoPR Contributes to Staphylococcus aureus Growth during Phosphate Starvation and Pathogenesis in an Environment-Specific Manner
    Article Snippet: .. Fragments were cloned into the pKOR1 knockout vector via site-specific recombination using the Gateway BP Clonase II enzyme mix (Thermo Fisher Scientific). .. The deletions were constructed via allelic exchange in Newman and the Δ pstSCAB Δ nptA mutant ( ) , as described previously ( ).

    Polymerase Chain Reaction:

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes
    Article Snippet: .. 150 ng/µl entry vector (e.g., pDONR™221; Life Technologies) Gateway® BP Clonase® II enzyme mix (Life Technologies) 10 to 50 ng/µl att B-PCR product prepared according to the Gateway® scheme (Basic Protocol 1) PCR-grade H2 O LB plates supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) LB medium supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) 50% (v/v) glycerol, sterile Plasmid DNA isolation kit (e.g., Qiagen; optional) 96-well plate or 1.5 ml microcentrifuge tubes Multichannel pipettor 25°C incubator Additional reagents and equipment for growth of T1 phage resistant (T1R ) E. coli competent cells (e.g. One Shot® MAX Efficiency® DH5α™ competent cells; Life Technologies) ( unit 1.8 ), DNA miniprep ( unit 1.6 or commercial DNA prep kit), and DNA sequencing (Chapter 7). ..

    Article Title: Metabolic Characterization of the Anthocyanidin Reductase Pathway Involved in the Biosynthesis of Flavan-3-ols in Elite Shuchazao Tea (Camellia sinensis) Cultivar in the Field
    Article Snippet: .. The resulting PCR products were purified and then cloned to the entry vector pDONR207 with Gateway® BP Clonase® Enzyme mix according to the manufacturer’s instructions (Invitrogen). .. The resulting ligation products were introduced to competent DH5α cells, which were selected on agar-solidified LB plates containing gentamycin.

    Plasmid Preparation:

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes
    Article Snippet: .. 150 ng/µl entry vector (e.g., pDONR™221; Life Technologies) Gateway® BP Clonase® II enzyme mix (Life Technologies) 10 to 50 ng/µl att B-PCR product prepared according to the Gateway® scheme (Basic Protocol 1) PCR-grade H2 O LB plates supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) LB medium supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) 50% (v/v) glycerol, sterile Plasmid DNA isolation kit (e.g., Qiagen; optional) 96-well plate or 1.5 ml microcentrifuge tubes Multichannel pipettor 25°C incubator Additional reagents and equipment for growth of T1 phage resistant (T1R ) E. coli competent cells (e.g. One Shot® MAX Efficiency® DH5α™ competent cells; Life Technologies) ( unit 1.8 ), DNA miniprep ( unit 1.6 or commercial DNA prep kit), and DNA sequencing (Chapter 7). ..

    Article Title: Metabolic Characterization of the Anthocyanidin Reductase Pathway Involved in the Biosynthesis of Flavan-3-ols in Elite Shuchazao Tea (Camellia sinensis) Cultivar in the Field
    Article Snippet: .. The resulting PCR products were purified and then cloned to the entry vector pDONR207 with Gateway® BP Clonase® Enzyme mix according to the manufacturer’s instructions (Invitrogen). .. The resulting ligation products were introduced to competent DH5α cells, which were selected on agar-solidified LB plates containing gentamycin.

    Article Title: Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT)
    Article Snippet: .. Purified cDNA ligated with three different forward adapters were mixed and recombined with the pDONR™ 222 plasmid vector via catalysis, using the Gateway® BP Clonase™ II Enzyme Mix (Invitrogen). ..

    Article Title: PhoPR Contributes to Staphylococcus aureus Growth during Phosphate Starvation and Pathogenesis in an Environment-Specific Manner
    Article Snippet: .. Fragments were cloned into the pKOR1 knockout vector via site-specific recombination using the Gateway BP Clonase II enzyme mix (Thermo Fisher Scientific). .. The deletions were constructed via allelic exchange in Newman and the Δ pstSCAB Δ nptA mutant ( ) , as described previously ( ).

    DNA Sequencing:

    Article Title: Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes
    Article Snippet: .. 150 ng/µl entry vector (e.g., pDONR™221; Life Technologies) Gateway® BP Clonase® II enzyme mix (Life Technologies) 10 to 50 ng/µl att B-PCR product prepared according to the Gateway® scheme (Basic Protocol 1) PCR-grade H2 O LB plates supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) LB medium supplemented with 50 µg/ml kanamycin (or other appropriate antibiotic; unit 1.1 ) 50% (v/v) glycerol, sterile Plasmid DNA isolation kit (e.g., Qiagen; optional) 96-well plate or 1.5 ml microcentrifuge tubes Multichannel pipettor 25°C incubator Additional reagents and equipment for growth of T1 phage resistant (T1R ) E. coli competent cells (e.g. One Shot® MAX Efficiency® DH5α™ competent cells; Life Technologies) ( unit 1.8 ), DNA miniprep ( unit 1.6 or commercial DNA prep kit), and DNA sequencing (Chapter 7). ..

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  • 99
    Thermo Fisher gateway bp clonase ii enzyme mix
    Gateway Bp Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 438 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway bp clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 438 article reviews
    Price from $9.99 to $1999.99
    gateway bp clonase ii enzyme mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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